Integrated network pharmacology, in vivo experiment, computational biology, and metabolomics to explore the mechanism of astragaloside IV against spinal cord injury

Spinal cord injury has become a global public health challenge. Astragaloside IV has been shown to have neuroprotective effects. However, the underlying mechanism of astragaloside IV against spinal cord injury remains poorly understood.

An integrated approach combining network pharmacology, in vivo experimental validation, molecular docking, molecular dynamics simulation, and metabolomics was used to investigate the underlying mechanism of astragaloside IV against spinal cord injury. Network pharmacology results indicated that IL6, IL1B, AKT1, TNF, and PTGS2 might be the key targets, while the TNF signaling pathway, IL17 signaling pathway, and C-type lectin receptor signaling pathway might be the critical signaling pathways involved in the mechanism of astragaloside IV against spinal cord injury. In vivo studies indicated that astragaloside IV significantly improved BBB scores and Rivlin inclined plane test performance in spinal cord injury mouse models, while reducing TNF-α, IL1B, IL6, and PTGS2 levels and increasing AKT1 expression.

Molecular docking results revealed that astragaloside IV exhibits strong binding affinity with IL1B, IL6, PTGS2, and AKT1. Molecular dynamics simulation further indicated that astragaloside IV could form stable complexes with AKT1 with a binding free energy of -27.17±3.21 kcal/mol. Metabolomic analysis further elucidated that the mechanism of astragaloside IV against spinal cord injury might involve up-regulating energy metabolism, nucleotide metabolism, amino acid metabolism, etc., and down-regulating lipid metabolism including linoleic acid metabolism, fatty acid biosynthesis, and α-linolenic acid metabolism.

Astragaloside IV can improve spinal cord injury through multiple targets, multiple signaling pathways, and multiple metabolic pathways, providing a new perspective on the molecular and metabolic mechanism network of spinal cord injury and strong experimental evidence for the use of astragaloside IV in treating spinal cord injury. RAC-alpha serine/threonine-protein kinase ‌Molecular mechanics generalized born surface area Partial least squares discriminant analysis Orthogonal partial least squares discriminant analysis This publication was funded by the Natural Science Foundation of Hunan Province (No. 2023JJ60022 & No. 2024JJ7568), Joint Funding Project of Hunan Provincial Natural Science Foundation and Hunan Xiangya Boai Rehabilitation Hospital Co Ltd. (No. 2026JJ81573) and China Postdoctoral Science Foundation (No. 2025M782316). Department of Spine Surgery, Xiangya Hospital of Central South University, Changsha, China Hunan Engineering Laboratory of Advanced Artificial Osteo-materials, Xiangya Hospital of Central South University, Changsha, China National Clinical Research Center for Geriatric Disease (Xiangya), Xiangya Hospital of Central South University, Changsha, China Department of Spine Surgery, Yiyang Central Hospital, Yiyang, China Department of Spine Surgery, Hunan Provincial People’s Hospital and The First-affiliated Hospital of Hunan Normal University, Changsha, China Correspondence to Xiaoyang Pang or Xiongjie Shen.

The authors declare no competing interests. This study was approved by the Experimental Animal Ethics Committee of Xiangya Hospital of Central South University (No.202407124). The authors have designated Xiaoyang Pang and Xiongjie Shen as corresponding authors who will communicate with the journal during the editorial process.

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